MYB转录因子在药用植物次生代谢领域的研究进展

MYB转录因子是植物中最大的转录因子家族之一,其生物学功能广泛,除去参与植物生长发育、生理活动代谢、细胞形态及模式建成、初生和次生代谢反应、对生物和非生物胁迫的应答等众多生物学过程外,在药用植物的次生代谢方面也颇具影响。目前关于MYB转录因子的研究多集中于农作物上,对于药用植物的研究少之又少,仍有很大研究空间。本研究综述了植物MYB转录因子的进化史、特征分类、功能及转化机制,着重探讨了MYB转录因子在植物次生代谢方面所起的作用,归纳总结前人相关的研究方法与思路,阐述国内外相关研究MYB的进展,为从多层面深入研究及更好地利用药用植物MYB转录因子提供思路和方法。     

绿竹咖啡酰辅酶A-O-甲基转移酶基因的克隆与分析

咖啡酰辅酶A-O-甲基转移酶（CCoAOMT）是木质素生物合成过程中的一个关键酶类。采用RT-PCR及RACE方法从绿竹中分离了CCoAOMT家族的一个基因,命名为BoCCoAOMT1（GenBank注册号为EF028662）。BoCCoAOMT1的cDNA全长873bp,含有一个780bp的读码框,编码一个259aa的蛋白,分子量约为29kD。序列分析结果表明,BoCCoAOMT1的编码肽链含有CCoAOMT类基因所有的保守序列元件,与禾本科植物水稻的CCoAOMT类基因有着很高的一致性,达到90.0%。系统进化树分析表明,BoC-CoAOMT1与水稻的CCoAOMT亲缘关系较近。     

香蕉MuMA DS2的克隆、序列分析和表达分析

根据抑制缩减文库获得的一个MADS-box基因片段,从香蕉果实cDNA文库中,采用RT-PCR技术分离获得一个全长cDNA,经序列测定和同源性分析表明,该cDNA含705bp的完整的开放阅读框,编码234个氨基酸残基的MADS-box蛋白,将其命名为MuMADS2。MuMADS2具有植物MADS-box基因家族中特有的M、I、K和C四个结构域,与芦笋、风信子、蝴蝶兰和百合的MADS-box基因的氨基酸序列存在很高的一致性（82%、78%、77%和75%）。将MuMADS2与其它植物来源的MADS-box基因进行序列比较发现,MuMADS2属于MADS-box基因的AGAMOUS亚族的D世系。RT-PCR分析表明,MuMADS2仅在生殖器官（花和果）中表达,在营养器官（根、茎和叶）中不表达。进一步分析发现MuMADS2仅在雄花和雌花的子房、由雌花发育而成的果实中表达,而在雄花和和雌花的柱头和花柱都不表达。推测MuMADS2可能在果实的发育和成熟过程中起作用。     

Metabolic characterization of tomato fruit during preharvest development, ripening, and postharvest shelf-life

Tomato is an important crop in terms of its economic and nutritional value. Tomato fruit quality is a function of metabolite content, which is prone to physiological changes related to fruit development and ripening. The aim of this work was to use a metabolomic approach to characterize compositional changes (sugars, acids and amino acids) of tomato during preharvest fruit development, ripening, and postharvest shelf-life. Gas chromatography-mass spectrometry was used to identify both polar and volatile metabolites that were involved in fruit development and ripening. Characteristic metabolites for the various fruit developmental stages were identified. Mannose, citramalic, gluconic and keto-l-gulonic acids were shown to be strongly correlated to final postharvest stages. During on-vine ripening, an increase was observed for the major hexoses, glucose and fructose, cell wall components such as galacturonic acid, and for amino acids such as aspartic, glutamic acid and methionine. Major changes were also observed at the level of the TCA cycle, showing a decrease in malic and fumaric acids, and accumulation of citric acid.     

Transformation of Diploid and Tetraploid Potato Clones with the rol C Gene of Agrobacterium rhizogenes and Characterization of Transgenic Plants

Diploid and tetraploid potato clones were transformed with the rol C gene from Agrobacterium rhizogenes under the control of the cauliflower-35-S promoter. Parameters proper to the sink and source attributes and hormone treatments have been considered to study the effect of the gene on yield physiology of the potato plant. The expression of the rol C gene in diploid and tetraploid potato lines induced drastic developmental abnormalities like dwarfism and increased tillering. Hormonal treatments revealed a different behavior of rol C transformams and controls. This supports the conclusion that the gene product of the ml C gene interacts with the hormonal status of the plant. Grafting experiments showed the non-ditfusable nature of rol C gene product. Tuberization was studied in-vitro and in-planta. Relevant differences in tuber shape and eye number were noted between rol C transfonnams and controls. Tuber yields in-planta in greenhouse or in growth chamber resulted higher in the controls; a significant increased number of tubers per plant was found, however, in both diploid and tetraploid rol C transformants. The effect of the rol C gene was apparently too drastic for diploid and tetraploid potatoes; weaker and tissue specific promoters may be necessary to induce only those modifications which are wanted for this crop. The experiments nevertheless proved that one single gene was capable in potato to affect substantially the manifestation of vieldmg attributes.     

转基因小麦目标基因通过花粉漂流的可能性研究

以去雄小麦为受体,在10m范围内小麦花粉可在任何方向发生有效漂移（使受体结实）,花粉的最远有效漂移距离可达80m,以不去雄的小麦、柱穗山羊草和卵穗山羊草为受体时,在距花粉源1m的范围内,小麦间的异交率最高为0.24%、小麦与柱穗山羊草和卵穗山羊草间的异交率分别为0.25%和0,花粉的最远有效漂移距离可达20m。依照欧盟的标准,非转基因小麦即使与转基因品种相邻种植,其产品中的转基因成分也不会超标,在麦类近缘野生植物的起源中心及主要分布区,释放转基因小麦要特别慎重。     

蒙古冰草Lea3抗旱基因片段的分离与鉴定

研究旨在利用同源序列法分离蒙古冰革抗旱基因同源片段。根据已知禾本科植物抗旱基因Lea第3组的保守区段设计一对简并引物,采用PCR方法对蒙古冰草幼苗的基因组DNA进行扩增。获得了1个蒙古冰草的抗旱基因Lea3基因片段,长度为497bp,包含124bp的非翻译区,共编码124个氨基酸。核苷酸序列分析表明,该序列与小麦抗旱基因LEA第3组的Wrad19同源性为93％,与小麦受ABA诱导的WRAB1基因同源性为93％;与一个来源于玉米mRNA的逆境胁迫基因克隆9124同源性为96％;与大麦受ABA诱导的pHVA1基因同源性为94％;与大麦HVA1基因同源性为91％。Southern杂交表明该基因在蒙古冰草基因组中以基因家族形式存在。     

松墨天牛肌球蛋白轻链2基因的鉴定及表达模式

为阐明松墨天牛肌肉收缩机理和相关基因表达模式,用构建c DNA文库方法,从松墨天牛c DNA中克隆到松墨天牛肌球蛋白轻链2基因,命名为Ma MLC-2(Gen Bank:KM371003)。该基因c DNA为882 bp,其中开放阅读框(ORF)为618 bp,编码205个氨基酸,推测的蛋白等电点为4.66,分子量为22.26 k Da。Ma MLC-2的氨基酸序列与赤拟谷盗同源性最高,为96%,与东方蝼蛄等8种昆虫同源性为85%~88%。松墨天牛与赤拟谷盗处在系统发育树的同一分支上。Ma MLC-2属于亲水性氨基酸,存在2个EF-hand结构域,长度均为29个氨基酸,其中第1个EF-hand结构域含有Ca2+结合位点。SWISS-MODEL预测结果显示,Ma MLC-2有2个螺旋-环-螺旋结构。用RT-q PCR分析了Ma MLC-2基因的相对表达量,结果显示:蛹和幼虫的表达量高于成虫;成虫中足表达量最高,腹部表达量最低,各部位均有表达,且有显著差异(P0.05)。     

辣椒金属伴侣蛋白基因CaHPP7提高植物对铜和热胁迫的抗性

金属伴侣蛋白在植物抵抗重金属胁迫和低温、干旱、高盐等非生物胁迫中发挥重要作用,但其应对热胁迫时发挥作用的报道还很少.前期研究从辣椒中分离到一个能够响应热胁迫的金属伴侣蛋白基因CaHPP7,通过生物信息学和定量分析发现该基因的氨基酸序列含有1个HMA结构域,且其表达受高温、Cu2+、Cd2+等重金属,以及ABA(Absc...     

从天冬-玉米的示范种植探讨"粮-药间套"增益模式

采用天冬单作、玉米单作和天冬-玉米套作3种种植模式,考查了整个生产过程中其主要生产资料和劳动力的投入以及产值等关键指标.对不同种植模式的经济效益对比分析结果表明,天冬-玉米套作比天冬单作增收21.8%,比玉米单作增收564.9%.天冬单作虽比玉米单作增收446.0%,但天冬种植三年后才能收获药材,没有短期收益.因此,3种模式中天冬-玉米套作的效果最佳.这一研究结果为种植业提供了一条"粮-药间套"的有效增益模式.     

Cloning,Characterization,and Gene Annotation of Cellulose Synthase Genes from Arabidopsis thaliana

The mechanistic basis of cellulose biosynthesis in plants has gained ground during last decade or so.The isolation of plant eDNA clones encoding cotton homologs of the bacterial cellulose synthase catalytic subunit was a significant achievement,which promises the elucidation of cellulose biosynthesis.     

葡萄糖氧化酶基因转化棉花和抗性愈伤组织的获得

用葡萄糖氧化酶基因转化棉花 ,研究了影响转化和抗性愈伤组织获得的因素。结果表明 ,基因型和培养基构成是影响葡萄糖氧化酶基因转化棉花获得抗性愈伤组织的最关键因素 ,在本试验所采用的 8个基因型中 ,中棉所 1 3中 940 9(中棉所3 5)和中棉所 1 9较易诱导获得抗性愈伤组织 ,而珂字 2 0 1、中棉所 2 7等则较难 ;用于棉花遗传转化的感染时间以 7～ 1 0 min为宜 ,共培养时间以48h为宜 ;下胚轴比子叶易诱导获得抗性愈伤组织。在诱导愈伤组织的继代培养中 ,加入高浓度的卡那霉素是获得转化细胞的关键 ;用液体培养比用固体培养易筛选获得抗性转化愈伤组织     

杧果中DXS基因的鉴定及与其他植物的比较分析

1-脱氧-D-核酮糖5-磷酸合成酶(1-deoxy-D-xylulose-5-phosphate synthase, DXS)是2-C-甲基-D-赤藓糖醇-4-磷酸(2-C-Methyl-D-erythritol-4-phosphate, MEP)途径的第一个关键酶。本研究鉴定了杧果DXS基因,以拟南芥DXS基因为参考,通过BLASTP方法获得了番木瓜、金钱橘、甜橙、苹果、桃子、白梨、葡萄、中华猕猴桃、无油樟、小果野蕉的DXS基因的蛋白序列,共32条序列,对其进行了保守结构域、保守基序、系统发育、理化性质、蛋白二级结构分析等研究。保守结构域及保守基序分析结果表明,杧果DXS蛋白包含一个PLN02582结构域,保守结构域内包含6个motif,其宽度、排列顺序与大多数植物相同,说明DXS基因家族具有高度保守性。系统发育分析结果显示,单子叶植物纲与双子叶植物纲的DXS基因没有分别形成分支,推测在进化过程中DXS基因的分化在双子叶植物纲和单子叶植物纲形成之后。理化性质分析及蛋白二级结构分析结果显示,稳定性蛋白占65.63%,碱性蛋白占25%,所有蛋白均为亲水性蛋白。杧果DXS蛋白为酸性、不稳定蛋白,具有亲水性。杧果DXS蛋白的二级结构主要结构元件是无规则卷曲、α-螺旋。本研究为进一步阐明杧果DXS基因在萜类合成途径中的功能提供了理论依据。     

移栽Bt棉的生长发育及其碳氮代谢研究

2002-2003年在江苏高肥力棉田,移栽条件下对9个Bt基因棉花品种(系)的生长发育特点及其碳氮代谢特征进行了研究。结果表明,中棉所29等3个杂交种在整个生长期表现为棉株高增长快,叶面积增长量大,现蕾和成铃强度大,生殖器官干物重也高,叶片全氮、可溶性糖含量高,NR、GPT、Rubisco、Sucroase活性高,表现为生长发育两旺,常规Bt棉GK19和新棉33B盛花前株高、叶面积生长快,现蕾强度大,叶片全氮含量高、NR、GPT活性高,表现为前期生长旺盛;新洋822和苏抗103结铃盛期后叶面积生长快,现蕾和成铃强度大,碳氮代谢强,表现为后期生育旺盛;sGK321和鲁棉研16整个生育期氮代谢强度大,但可溶性糖含量低、Rubisco、Sucroase等活性小,营养生长快,生殖生长弱,表现为营养生长过旺。     

Growth and Metabolic Changes of Cowpea Plants as Affected by Water Deficiency and Indole-3-Yl-Acetic Acid

Cowpea plants ( Vigna sinensis Savi.) were grown in pots for two successive years. These plants were daily maintained water holding capacities (15, 25, 45, 65 %).
The foliage of plants grown at every level of soil moisture was sprayed till dripping with IAA solutions (0, 10, 50, 100 mg/l) three times after 3, 6 and 9 weeks from sowing. Samples were taken at the two physiological stages before flowering and during flowering for growth measurements and determinations of nitrogen fractions.
Water shortage led to retardation of stem length, number of internodes, number of leaves and dry weight shoots. IAA increasing the same parameters only when applied at concentrations of 10 and 50mg/1 at moisture level 65 %, 45 % and 25 % of W.H.C.
Water stress decreased total-N and protein-N contents of cowpea shoots. On the other hand total soluble- N content was increased under the same conditions. IAA increased total-N and protein-N content of cowpea shoots. Meanwhile, the same growth regulator decreased total soluble-N when applied at 45% 25 % and 15 % W.H.C. levels.     

Metabolic Engineering of Carotenoid Synthesis: Development of a Production System with Nicotiana glauca and Improvement of the Nutritional Value of Potato

Carotenoid pigments are of commercial interest as animal feed and play an important role in human metabolism. β-Carotene is important as provitamin A and lutein and zeaxanthin have to be constantly integrated into the macula lutea in the central retina of the eye to prevent age-dependent macular degeneration. Diets rich in carotenoids prevent the onset of certain chronic diseases and certain types of cancer.     

板栗叶水浸液对3种中药材种子萌发及幼苗生长的影响

选取商洛道地中药材桔梗、黄芩和丹参,研究不同浓度(0,5,10,20,30,40 mg/mL)板栗叶水浸液对3种受体中药材种子萌发和幼苗生长的影响.结果表明,低浓度板栗叶水浸液对3种中药材种子的萌发有明显的化感促进作用,但随着水浸液浓度的不断提高,其对受体种子的化感促进作用逐渐减弱并消失,开始表现为抑制作用.板栗叶水浸液在5～20 mg/mL浓度范围内对3种受体中药材种子的发芽率、发芽势、发芽指数、相对电导率、游离氨基酸含量、可溶性糖含量、可溶性蛋白含量以及幼苗根长、苗高均表现为化感促进作用,但超过其临界值就开始表现为化感抑制作用.其总体趋势呈“抛物线”形规律.板栗叶水浸液在试验浓度范围对桔梗、黄芩、丹参种子萌发和幼苗生长有一定的促进作用.     

甜瓜乙烯受体基因Cm-ETR2的克隆及其在果实发育成熟过程中表达特性的分析

为研究Cm-ETR2基因在甜瓜果实发育成熟过程中的作用机理,根据已报道的甜瓜(Cucumis melo)品种Cantalupensis的Cm-ETR2基因序列,设计特异性引物,通过RT-PCR技术从甜瓜品种河套蜜瓜(Cucumis melo L.cvHetao)成熟果实中克隆得到了乙烯受体基因Cm-ETR2 cDNA,序列分析显示,序列长度为2 304 bp,编码767个氨基酸。利用实时荧光定量RT-PCR方法,对其在河套蜜瓜果实发育成熟过程中的表达特性进行了分析,Cm-ETR2基因在15DAP至30DAP表达基本没有变化,35DAP表达量开始上升,上升趋势一直持续至45DAP,且45DAP表达量为15DAP的9倍。